The constructs pEGFP BIN1 full length (isoform 8, 454 aa) and pEGFP BAR+PI (aa 1-281) were obtained from P. De Camilli (Yale University, New York). cDNA corresponding to MTM1 sequence (603 aa) was cloned into pEGFP-C1 (Clontech, N-ter GFP fusion) by PCR amplification followed by BamH1 restriction and into pCS2+ vector ([1], untagged). cDNA corresponding to the full length, BAR+PI, BAR (aa 1-255...

BACKGROUND The main barriers to non-viral gene delivery include cellular and nuclear membranes. As such, the aim of this study was to develop a type of vector that can target cells through receptor-mediated pathways and by using nuclear localization signal (NLS) to increase the nuclear uptake of genetic materials. METHODS A dexamethasone (Dexa)-conjugated lipid was synthesized as the material...

OBJECTIVE To study the effects of hepatitis C virus gene nonstructural protein 2 on the expressions of Bcl-2 and Bax in liver hepatocellular cells. METHOD The study was conducted at the Department of Infectious Diseases, the First Affiliated Hospital of Medical School, Xi'an Jiaotong University, Xi'an, China, from March 2012 to April 2013. Negative controls pEGFP-C3-NS2, pEGFP-C3-C and pEGFP-...

Intermediate filament (IF) overproduction induces abnormal accumulation of neuronal IF, which is a pathological indicator of some neurodegenerative disorders. In our study, α-Internexin- and peripherin-overexpressing PC12 cells (pINT-EGFP and pEGFP-peripherin) were used as models to study neuropathological pathways responsible for neurodegenerative diseases. Microarray data revealed that Cdk5-r...

BACKGROUND Nanostructured lipid carriers (NLC), composed of solid and liquid lipids, and surfactants are potentially good colloidal drug carriers. The aim of this study was to develop surface-modified NLC as multifunctional nanomedicine for codelivery of enhanced green fluorescence protein plasmid (pEGFP) and doxorubicin (DOX). METHODS TWO DIFFERENT NANOCARRIERS: pEGFP- and DOX-loaded NLC, an...

The organic anion transporter, rROAT1, is a dicarboxylate/organic anion exchanger, a function associated with the basolateral membrane in rat proximal tubule. To directly establish the subcellular localization of rROAT1 in renal epithelia, we made a rROAT1-green fluorescent protein (GFP) fusion construct (rROAT1-GFP). Plasma membrane-associated fluorescence was observed in rROAT1-GFP-expressing...

A recombinant vesicular stomatitis virus (VSV-PeGFP-M-MmRFP) encoding enhanced green fluorescent protein fused in frame with P (PeGFP) in place of P and a fusion matrix protein (monomeric red fluorescent protein fused in frame at the carboxy terminus of M [MmRFP]) at the G-L gene junction, in addition to wild-type (wt) M protein in its normal location, was recovered, but the MmRFP was not incor...

The phosphoprotein (P) of vesicular stomatitis virus (VSV) is a subunit of the viral RNA polymerase. In previous studies, we demonstrated that insertion of 19 amino acids in the hinge region of the protein had no significant effect on P protein function. In the present study, we inserted full-length enhanced green fluorescent protein (eGFP) in frame into the hinge region of P and show that the ...

Immunological alterations have been reported to be involved in glioma, the most common malignant disease of the adult brain. Our recent study identified higher levels of IL-17 in glioma specimens. The present study investigated the role and possible mechanisms of IL-17 in glioma tumorigenesis. Human IL-17 cDNA was cloned and inserted into the eukaryotic pEGFP-N1 expression vector, which was use...

The mechanisms whereby mutant gene expression triggers neurodegeneration are poorly understood but have generally been attributed to translated gene products. We now demonstrate direct neuropathic effects of untranslated RNA on cultured motor neurons. We show that expression of untranslated light neurofilament (NF-L) RNA sequence in the 3'UTR of an EGFP transgene (pEGFP/NF-L RNA) or in a separa...